用于瞬时蛋白表达的HEK293细胞的高通量转染

High Throughput Transfection of HEK293 Cells for Transient Protein Production.

作者信息

Arena Tia A, Harms Peter D, Wong Athena W

机构信息

Department of Early Stage Cell Culture, Genentech Inc., South San Francisco, CA, USA.

Department of Late Stage Cell Culture, Genentech Inc., South San Francisco, CA, USA.

出版信息

Methods Mol Biol. 2018;1850:179-187. doi: 10.1007/978-1-4939-8730-6_12.

DOI:10.1007/978-1-4939-8730-6_12

PMID:30242687

Abstract

Transient transfection of mammalian cells is used in the biotechnology industry to quickly supply recombinant protein for research and large molecule drug development. Here, we describe a method for high throughput transient transfection of Human Embryonic Kidney 293 (HEK293) cells in 30 mL tubespins using polyethylenimine (PEI) as a transfection reagent. An automated liquid handler can be used to perform pipetting steps for transfecting batches of 96 tubespins, and septa in the tubespin caps allow for rapid processing without decapping. The addition of valproic acid (VPA) to transfection cultures enhances recombinant protein production. The thawing and passaging operations for HEK293 cultures to source the transient transfections are also described.

摘要

在生物技术产业中，哺乳动物细胞的瞬时转染用于快速提供重组蛋白，以用于研究和大分子药物开发。在此，我们描述了一种使用聚乙烯亚胺（PEI）作为转染试剂，在30 mL离心管中对人胚肾293（HEK293）细胞进行高通量瞬时转染的方法。可使用自动液体处理仪对96个离心管批次进行移液操作以进行转染，离心管帽中的隔片允许在不打开管帽的情况下快速处理。在转染培养物中添加丙戊酸（VPA）可提高重组蛋白产量。还描述了用于瞬时转染的HEK293培养物的解冻和传代操作。